AI Article Synopsis

  • PC is a crucial plasma anticoagulant, and mutations in both alleles can lead to a serious condition called neonatal purpura fulminans.
  • The study developed a genome editing approach using engineered activated protein C (APC) to treat congenital PC deficiency by expressing it in mouse liver through adeno-associated virus vectors and CRISPR/Cas9.
  • Results showed that the engineered APC prolonged coagulation time, inhibited harmful thrombus formation, and improved the survival of PC-deficient mice, indicating its potential as a cure for this severe condition.

Article Abstract

Background: PC (protein C) is a plasma anticoagulant encoded by ; mutation in both alleles results in neonatal purpura fulminans-a fatal systemic thrombotic disorder. In the present study, we aimed to develop a genome editing treatment to cure congenital PC deficiency.

Methods: We generated an engineered APC (activated PC) to insert a furin-cleaving peptide sequence between light and heavy chains. The engineered PC was expressed in the liver of mice using an adeno-associated virus vector or CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat-associated 9)-mediated genome editing using an adeno-associated virus vector in vivo.

Results: The engineered PC could be released in its activated form and significantly prolonged the plasma coagulation time independent of the cofactor activity of PS (protein S) in vitro. The adeno-associated virus vector-mediated expression of the engineered PC, but not wild-type PC, prolonged coagulation time owing to the inhibition of activated coagulation FV (factor V) in a dose-dependent manner and abolished pathological thrombus formation in vivo in C57BL/6J mice. The insertion of (enhanced green fluorescent protein) sequence conjugated with self-cleaving peptide sequence at locus via neonatal in vivo genome editing using adeno-associated virus vector resulted in the expression of EGFP in 7% of liver cells, mainly via homology-directed repair, in mice. Finally, we succeeded in improving the survival of PC-deficient mice by expressing the engineered PC via neonatal genome editing in vivo.

Conclusions: These results suggest that the expression of engineered PC via neonatal genome editing is a potential cure for severe congenital PC deficiency.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11594008PMC
http://dx.doi.org/10.1161/ATVBAHA.123.319460DOI Listing

Publication Analysis

Top Keywords

genome editing
24
adeno-associated virus
16
expression engineered
12
neonatal genome
12
virus vector
12
cure congenital
8
peptide sequence
8
regularly interspaced
8
interspaced short
8
short palindromic
8

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!